A Technique to Induce Chronic Experimental Autoimmune Dry Eye in Rats

Abstract

Source: Hou, A., et al. A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue. J. Vis. Exp. (2017).

This video demonstrates a technique to induce chronic experimental autoimmune dry eye disease in rats. Upon injecting an antigen-adjuvant emulsion containing autoantigens and foreign antigens into a rat at predefined intervals, chronic eye inflammation is induced in the immunized rat by injecting the same antigens into the ocular surface tissues.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation of Lacrimal Gland Extract NOTE: Rats were anesthetized with the intraperitoneal injection of ketamine (75 mg/kg) and xylazine (10 mg/kg). Proper anesthetization was confirmed by toe pinching and tail pinching. Ophthalmic gel was applied to the rat's eyes to prevent dryness …

Materials

Reagents
Protease inhibitor cocktail	Sigma-Aldrich	P2714-1BTL
Pierce BCA Protein Assay Kit	Thermal Scientific	23227
Mycobacterium tuberculosis H37Ra	Becton, Dickinson and company	231141
complete Freund's adjuvant	Becton, Dickinson and company	231131
ovalbumin	Sigma-Aldrich	A5503-10G
incomplete Freund's adjuvant	Sigma-Aldrich	F5506-6X10ML
pertussis toxin	Sigma-Aldrich	P7208-50UG
fluorescein sodium solution	Bausch & Lomb U.K Limited	NA
Equipment
Sonicator	Sonics	Vibra-Cell
Micro IV microscope	Phoenix Research Labs	NA